Automated Microfluidic Cell Separator

Project Group: 16083

The P16083 Group

Member Major Role Contact

Jay Dolas BME Lead Engineer jrd8174@rit.edu

Alexandra la Londe BME Microfludics Specialist ael9930@rit.edu

Vincent Serianni II BME Project Manager vcs7133@rit.edu

Tyler Lisec ME Lead Mechanical tjl9229@rit.edu

Ryan Kinney EE Lead Electrical rhk9387@rit.edu

Chris Molinari EE Controls Engineer cxm4025@rit.edu

“Microfludics is the field that studies the manipulation of small amounts of fluids (10-

9 to 10-18 liters)”

“Microfluidics refers to the handling of liquids or gases at a scale generally below 1mm, where a number of phenomena that are NOT present or not predominant at larger scales can be exploited for numerous purposes”.

“The field of microfluidics is in essence multidisciplinary as it combines microfabrication tecniques with chemistry and biology”.

BIME 489-01 Dr. Blanca Lapizco-Encinas

Background Summary – What is Microfluidics?

Background Summary – What is Dielectrophoretics?

Dielectrophoresis is the movement of particles due to polarization effects in a non-uniform electric field.

In the presence on a nonuniform electric field, one side of the dipole will be in a region with a lower field intensity.

This will produce and UNEVEN charge alignment in the particle, inducing it to move toward the regions of greater field strength

BIME 489-01 Dr. Blanca Lapizco-Encinas

http://cnx.org/resources/7b4d03a7fc1e724f75950258ae6d2356

Background Summary - What Size of Particles?

Research and Testing Teaching Tool General Medical

Purposes

http://www.mdpi.com/1422-0067/15/10/18281/htm

Background Summary – Why Sort Cells?

http://eshop.eppendorf.ca/upload/productView/Eppendorf_5427R_high-capacity_centrifuge.jpg

Background Summary - Other Sorting Methods

Centrifuge Flow Cytometry Channel Geometry Magnetic Based

A cell separator is a device that separates cells in a mixture, based upon pre-established criteria (biomarkers, size, electrical characteristics, etc.). This is necessary in many cell culture and diagnostic applications where downstream processes occur after cell culture, such as purification or analysis. Optimally, this device should not interfere with the viability or characteristics of the cells, while still being cost effective. Current cell separation devices require some sort of labeling (either fluorescent or magnetic) which is not only costly but can affect cell behavior and mortality. We propose an automated microfluidic system that utilizes developing technologies (dielectrophoretics) to reduce costs drastically while maintaining cell viability.

The goals of this project are to develop a system that not only sorts cells without the use of labeling, but also fits within a biosafety cabinet, is self-driven, and is automated (hands-off once the sample is loaded and sequence has started). The expected result is a functional prototype that fits all of the goals above and is suitable for use in a teaching laboratory. The design and prototype must conform to intellectual property and diagnostic laboratory standards so that it may be marketed this as a definitive step forwards in cell separation technology.

Project Statement

http://www.appliedcytometry.com/flow_cytometry.php

http://www.elveflow.com/microfluidic-tutorials/cell-biology-imaging-reviews-and-tutorials/microfluidic-for-cell-biology/label-free-microfluidic-cell-separation-and-sorting-techniques-a-review/

Current State

Flow Cytometry Fluorescence labeling

Laser to excite and identify the cells

Additives could alter or damage cells

Hydrodynamic Cell Separator Inertial forces to separate cell types

High shear forces can damage cells

Dielectrophoresis Uses electric fields to

manipulate the cell location in a stream

No need for additives

No added shear force

http://www.elveflow.com/microfluidic-tutorials/cell-biology-imaging-reviews-and-tutorials/microfluidic-for-cell-biology/label-free-microfluidic-cell-separation-and-sorting-techniques-a-review/

Desired State

Working prototype that can: Separate cells to demonstrate, in a class setting, the use of

dielectrophoretics in cell separation

Act as a partially automated system

User only has to load the sample and set the target specifications

Maintain cell viability during sorting process

Accurately sorts the target cells

Documentation of the prototype that illustrates: Proper use and care of the device

Target specifications for certain cells

Voltage amplitude and frequency standards in order to sort a given cell

Project Goals and Deliverables

Device start-up cost

Weight, to be moved by 1 person

Electrical shielding and insulation

Bio-hazard containment

Footprint (2' x 1')

120V outlet compatible

Reusable channel

Channel is visible under microscope during run

Preform process within one hour

Key Constraints

Use Scenarios – Teaching Aid

Use Scenarios – Medical Field

http://www.rheonix.com/corporate/careers.phphttps://twitter.com/ritbme

Stakeholders

Customer – Dr. Blanca Lapizco-Encinas End Users – Lab Workers, Professors, Researchers, Students Potential Sponsors – Rheonix Inc. or the BME Department at

RIT Other Stakeholders – P16083 Group, MSD Team

Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important

Customer Requirements

Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important

Customer Requirements

Separation can be visualized

Engineering Requirements

Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important

Engineering Requirements

Scale: 1 = Less Important, 3 = Moderately Important, 9 = Very Important

n = Less Importantn = Moderate Importancen = Most Important

House of Quality

Project Plans

MSD I: Introduced to the project and group

Gather data for design and hardware

Research, Design, Review, Standards, Bill of Materials (BOM)

Interact with the Customer

Interview, Funding, Customer vs Engineering Requirements

MSD II: Build validated designs

Circuits, PDMS Channels, 3D Print (where needed), Subsystems, Benchmarking

Present project in working state

Imagine RIT, Class Demo

Project Plans (First 3 Weeks)

Project Plans (Next 3 Weeks)

Project Plans (MSD I)

Risks

http://www.clker.com/cliparts/O/c/x/E/f/v/fire-hazard-md.png

Technical

Power Supply Failure

Size/Weight

Channel Fabrication Errors

Fluid Flow Pressure

Cells Having Similar Electrical Properties

Cell Viability

http://images.clipartpanda.com/no-money-sign-clip-art-money-sign.png

Resource

Expense of Components

Lack of Manpower

Data Gathering

Cells for sorting

Clean Room Workers

http://www.health-safety-signs.uk.com/productimages/Danger-Electric-shock-risk-sign.gif

http://www.emedco.com/media/catalog/product/Indl-Eyewash-Shower--First-Aid-Signs-42878BBVPLY2WY-ba.jpg

Safety

Electric Shock

Lab Work

Channel Fabrication

Leaks

http://cdn2.bigcommerce.com/server4600/10c6f/products/1621/images/3022/WS32001_warning_airborne_toxic_chemicals__69205.1403023543.1280.1280.png?c=2

Environmental & Societal

Waste Generation

Potential Air Born Chemicals

Disposal of Chemicals

Class Room Setting